JP7403575B2 - Method of reducing serum levels of Fc-containing agents using FcRn antagonists - Google Patents

Description

(Related application)
This application claims the benefit of U.S. Provisional Application No. 62/130,076, filed March 9, 2015, which is incorporated herein by reference in its entirety.

(background)
Immunoglobulin gamma (IgG) antibodies play a role in the pathology of many disorders, such as autoimmune diseases, inflammatory diseases, and disorders whose pathology is characterized by overexpression of IgG antibodies (e.g., hypergammaglobulinemia). (e.g. Junghans literature, Immunologic Research
16(1):29 (1997)).

The half-life of IgG in serum is long compared to the serum half-life of other plasma proteins (Roope
Nian et al., J. Immunology 170:3528 (2003); Junghans and Anderson, Proc. N
atl. Acad. Sci. USA 93:5512 (1996)). This long half-life is due, in part, to the binding of the Fc region of IgG to the Fc receptor, FcRn. Although FcRn was initially characterized as a fetal transport receptor for maternally inherited IgG, FcRn also functions to protect the IgG from degradation in the adult. FcRn binds pinocytically taken up IgG and recirculates it back to the extracellular compartment, thereby removing it from transport to lysosomes where it is responsible for degradation.
Protects IgG. This recycling is facilitated by pH-dependent binding of IgG to FcRn. Here, the IgG/FcRn interaction is stronger at acidic endosomal pH than at extracellular physiological pH.

When the serum concentration of IgG reaches a level that exceeds the available FcRn molecules, unbound IgG is no longer protected from degradation mechanisms and therefore has a reduced serum half-life. Therefore, inhibiting IgG binding to FcRn reduces the serum half-life of IgG by preventing IgG recycling by endosomes. Accordingly, agents that antagonize IgG binding to FcRn may be useful in modulating, treating, or preventing antibody-mediated disorders such as autoimmune diseases, inflammatory diseases, and the like. One example of a method to antagonize the binding of IgG Fc to FcRn involves the generation of blocking antibodies to FcRn (see, eg, WO2002/43658). In addition, peptides that bind to and antagonize the function of FcRn have also been identified (for example, US6,212,022
and US 8,101,186). Additionally, full-length IgG antibodies containing variant Fc receptors with enhanced FcRn binding and reduced pH dependence have also been identified to antagonize FcRn binding to IgG (see, e.g., 8,163,881). . However, there is a need in the art for improved methods of treating antibody-mediated disorders.

(overview)
The present disclosure provides novel methods of reducing serum levels of Fc-containing agents (eg, antibodies and immunoadhesins) in a subject. These methods generally target Fc native to the subject.
specifically binds FcRn with increased affinity and reduced pH dependence compared to the
comprising administering an effective amount of an isolated FcRn antagonist. The disclosed methods are particularly useful for treating antibody-mediated disorders, such as autoimmune diseases.

Accordingly, in one aspect, the present disclosure provides a method of reducing serum levels of an Fc-containing agent in a subject, comprising administering to the subject an effective amount of an isolated FcRn antagonist comprising a variant Fc region or an FcRn-binding fragment thereof. wherein the Fc domain of the variant Fc region is
amino acids Y, T, E, K, F, and Y at positions 252, 254, 256, 433, 434, and 436, respectively, and the FcRn antagonist is administered to the subject at a dose of about 0.2 to about 200 mg/kg. The above method is provided.

In another aspect, the present disclosure provides a method for reducing serum levels of an Fc-containing agent in a subject, the method comprising: administering to the subject an effective amount of an isolated
administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25.
2, 254, 256, 433, 434, and 436 each contain the amino acids Y, T, E, K, F, and Y, and the FcRn antagonist is administered to the subject at least twice every 20 days. I will provide a.

The following embodiments apply to all aspects of this disclosure.

In certain embodiments, the FcRn antagonist is administered to the subject 1, 2, 3, 4, 5, 6, 7, 8,
Administered once every 9 or 10 days. In certain embodiments, the FcRn antagonist is administered to the subject once every four days. In certain embodiments, the FcRn antagonist is administered to the subject once every 7 days. In certain embodiments, the FcRn antagonist is administered to the subject on day 20 at 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18
, 19, or 20 doses.

In another aspect, the present disclosure provides a method of reducing serum levels of an Fc-containing agent in a subject, the method comprising: administering to the subject an effective amount of an isolated
administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25.
2, 254, 256, 433, 434, and 436 contain the amino acids Y, T, E, K, F, and Y, respectively, and the FcRn antagonist is administered to the subject once every 48 hours for 4 weeks. The above method is provided.

In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 0.2 to about 200 mg/kg. In certain embodiments, the FcRn antagonist administers to the subject about 0.2, 1
, 2, 3, 5, 10, 20, 25, 30, 50, 70, 100, or 200 mg/kg. In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 10 mg/kg.
In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 20 mg/kg. In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 25 mg/kg.

In another aspect, the present disclosure provides a method of reducing serum levels of an Fc-containing agent in a subject, the method comprising: administering to the subject an effective amount of an isolated
administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25.
2, 254, 256, 433, 434, and 436 contain the amino acids Y, T, E, K, F, and Y, respectively, and the FcRn antagonist is administered to the subject once every 4 days at a dose of about 25 mg/kg. The method is provided in which the method is administered at a frequency of one or more times.

In another aspect, the present disclosure provides a method of reducing serum levels of an Fc-containing agent in a subject, the method comprising: administering to the subject an effective amount of an isolated
administering an FcRn antagonist, wherein the Fc domain of the variant Fc region is at EU position 25.
2, 254, 256, 433, 434, and 436 contain the amino acids Y, T, E, K, F, and Y, respectively, and the FcRn antagonist is administered to the subject once every 7 days at a dose of about 25 mg/kg. The method is provided in which the method is administered at a frequency of one or more times.

In certain embodiments, the FcRn antagonist is administered intravenously. In certain embodiments, the FcRn antagonist is administered subcutaneously. In certain embodiments, the FcRn antagonist is administered to the subject in two or more doses, wherein the first dose administered to the subject is administered intravenously and one or more of the second or subsequent doses is administered subcutaneously.

In certain embodiments, the FcRn antagonist does not include antibody variable regions. In certain embodiments, the FcRn antagonist does not include a CH1 domain. In certain embodiments, the FcRn antagonist does not contain free cysteine residues. In certain embodiments, the variant Fc region is an IgG Fc region. In certain embodiments, the variant Fc region is an IgG1 Fc region. In certain embodiments, the amino acid sequence of the Fc domain of the variant Fc region comprises the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3. In certain embodiments, the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO:1. In certain embodiments, the FcRn antagonist consists of a variant Fc region, and the amino acid sequence of the Fc domain of the variant Fc region is SEQ ID NO: 1, 2, or
It consists of the amino acid sequence described in 3.

In certain embodiments, the variant Fc region has increased affinity for Fcγ receptors compared to the affinity of a wild-type IgG1 Fc region for Fcγ receptors. In certain embodiments, the variant Fc region has increased affinity for CD16a. In certain embodiments, the Fc domain of the variant Fc region does not include an N-linked glycan at EU position 297. In certain embodiments, the FcRn antagonist comprises a plurality of FcRn antagonist molecules, and at least 50% (optionally, at least 60, 70, 80%) of the plurality of FcRn antagonist molecules.
, 90, 95, or 99%) contains an afucosylated N-linked glycan at EU position 297.
c region or its FcRn-binding fragment. In certain embodiments, the FcRn antagonist comprises a plurality of FcRn antagonist molecules, and at least 50 of the plurality of FcRn antagonist molecules
% (optionally at least 60, 70, 80, 90, 95, or 99%) contain a variant Fc region or FcRn binding fragment thereof that includes an N-linked glycan with a bisecting GlcNac at EU position 297.

In certain embodiments, the variant Fc region is linked to a half-life extender. In certain embodiments, the half-life extender is polyethylene glycol or human serum albumin.

In certain embodiments, the Fc-containing agent is an antibody or an immunoadhesin. In certain embodiments, the Fc-containing agent is a therapeutic or diagnostic agent. In certain embodiments, the
The Fc-containing agent is a contrast agent. In certain embodiments, the Fc-containing agent is an antibody drug conjugate. In certain embodiments, the Fc-containing agent is a pathogenic antibody. In certain embodiments, the Fc-containing agent is an autoantibody.

In certain embodiments, the subject has an antibody-mediated disease or disorder, wherein administration of the FcRn antagonist to the subject ameliorates the disease or disorder. In certain embodiments, the disease or disorder is treatable using intravenous immunoglobulin (IVIG), plasmapheresis, and/or immunoadsorption. In certain embodiments, the antibody-mediated disease or disorder is an autoimmune disease. In certain embodiments, the autoimmune disease is allogeneic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune Addison's disease, Alzheimer's disease, antineutrophil Cytoplasmic autoantibodies (ANCA), autoimmune diseases of the adrenal glands,
autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, autoimmune neutropenia,
Autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman syndrome, celiac disease-dermatitis (celia
c spruce-dermatitis), chronic fatigue immunodeficiency syndrome, and chronic inflammatory demyelinating polyneuritis (CIDP).
), Churg-Strauss syndrome, cicatrical pemphigoid, Crest syndrome, cold agglutinin disease, Crohn's disease, dermatomyositis, dilated cardiomyopathy, discoid lupus erythematosus, epidermolysis bullosa acquired, mixed essential type cryoglobulinemia, factor VIII deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barré, Goodpasture syndrome,
Graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic membranous neuropathy, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, IgM polyneuropathy, immunology Mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen plantus
, lichen moss sclerosus, lupus erthematosis, Meniere's disease, mixed connective tissue disease, mucosal pemphigoid, multiple sclerosis, type 1 diabetes, multifocal motor neuropathy (MMN), myasthenia gravis, paraneoplastic bullous pemphigoid, gestational pemphigoid, pemphigus vulgaris, pemphigus foliaceus,
Pernicious anemia, polyarteritis nodosa, polychrondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia
agammaglobinulinemia), primary biliary cirrhosis, psoriasis, psoriatic arthritis, relapsing polychondritis, Reynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, solid Organ transplant rejection, stiff man syndrome, systemic lupus erythematosus, Takayasu arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (SJS), temporal arteritis/giant cell arteritis giant cell arteritis), thrombotic thrombocytopenic purpura, ulcerative colitis, uveitis, dermatitis herpetiformis vasculitis
, anti-neutrophil cytoplasmic antibody-associated vasculitis, vitiligo, and Wegener's granulomatosis.

In certain embodiments, the autoimmune disease is an autoimmune channelopathy. In certain embodiments, the channelopathy is autoimmune limbic encephalitis, epilepsy, neuromyelitis optica,
Lambert-Eaton myasthenia syndrome, myasthenia gravis, anti-N-methyl-D-aspartate (N
MDA) receptor encephalitis, anti-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor encephalitis, Morvan syndrome, myotonia nervosa, streptococcal infection (PANDAS), and glycine receptor antibody-related disorders. In certain embodiments, the antibody-mediated disorder is hyperglobulinemia.

In certain embodiments, the FcRn antagonist is administered to the subject simultaneously or sequentially with an additional therapeutic agent. In certain embodiments, the additional therapeutic agent is an anti-inflammatory agent. In certain embodiments, the additional therapeutic agent is a leukapheresis agent. In certain embodiments, the leukocyte depleting agent is a B cell depleting agent. In certain embodiments, the B cell depleting agent is an antibody. In certain embodiments, the B cell depleting agent is CD10, CD19, CD20, CD2
1, CD22, CD23, CD24, CD37, CD53, CD70, CD72, CD74, CD75, CD77, CD79a, CD79b, CD8
0, an antibody that specifically binds to CD81, CD82, CD83, CD84, CD85, or CD86. In certain embodiments, the additional therapeutic agent is rituximab, daclizumab, basiliximab, muronomab-CD3, infliximab, adalimumab, omalizumab, efalizumab, natalizumab, tocilizumab, eculizumab, golimumab, canakinumab, ustekinumab, belimumab, or the like. It is a combination of

In certain embodiments, the subject is a human or a cynomolgus monkey.

(Brief explanation of the drawing)
Figure 1 shows the results of an experiment to determine the effect of Fc-Abdeg and HEL-Abdeg on serum levels of tracer antibody (FR70-hIgG1) in cynomolgous monkeys. Figure 2 shows the results of an experiment to determine the effect of Fc-Abdeg and HEL-Abdeg on serum levels of total IgG in cynomolgus monkeys. Figure 3 shows the results of an experiment to determine the effects of Fc-Abdeg and HEL-Abdeg on albumin levels in cynomolgus monkeys. Figure 4 shows the results of an experiment to determine the effect of Fc-Abdeg and IVIG on serum levels of tracer antibody (FR70-hIgG1) in cynomolgus monkeys. Figure 5 shows the results of an ELISA assay comparing the affinity of Fc-Abdeg, Fc-Abdeg-POT and Fc-Abdeg-S239D/I332E for human CD16a. Figure 6 shows the results of an ELISA assay comparing the affinity of Fc-Abdeg, Fc-Abdeg-POT and Fc-Abdeg-S239D/I332E for mouse CD16-2. Figure 7 shows the results of an experiment to determine the effects of Fc-Abdeg, Abdeg-POT and Fc-AbdegS239D/I332E on anti-CD20-induced ADCC signals using Promega's Raji-based ADCC reporter bioassay. There is. Figure 8 shows the results of an experiment to determine the effect of Fc-Abdeg and Abdeg-POT on anti-CD70-induced lysis of CD70+ U266 cells in vitro. Figure 9 shows the results of experiments to determine the effects of Fc-Abdeg, Fc-Abdeg-POT, Fc-Abdeg-S239D/I332E and IVIG on platelet levels in an acute mouse model of immune thrombocytopenia. Figure 10 shows the results of an exemplary gel filtration purification of Fc-Abdeg. Figure 11 shows the results of a dose escalation study measuring the effect of various single doses of Fc-Abdeg on serum levels of tracer antibody (FR70-hIgG1) in cynomolgus monkeys. Figure 12 shows the results of a dose escalation study measuring the effect of various single doses of Fc-Abdeg on serum levels of tracer antibody (FR70-hIgG1) in cynomolgus monkeys. Figure 13 shows the results of an experiment to determine the effect of 200 mg/kg Fc-Abdeg on the levels of cIgA in cynomolgus monkeys. Figure 14 shows the results of an experiment to determine the effect of 200 mg/kg Fc-Abdeg on the levels of cIgM in cynomolgus monkeys. Figure 15 shows the pharmacokinetic profile of various single doses of Fc-Abdeg in cynomolgus monkeys. Figure 16 shows the results of an experiment to determine the effect of repeated multiple doses of 20 mg/kg Fc-Abdeg on the levels of cIgG in cynomolgus monkeys. Figure 17 shows the pharmacokinetic profile of Fc-Abdeg in cynomolgus monkeys given repeated multiple doses of 20 mg/kg Fc-Abdeg. Figure 18 shows the results of a dose escalation study measuring the effect of various single doses of Fc-Abdeg on serum levels of endogenous IgG levels in cynomolgus monkeys. Figure 19 shows the pharmacokinetic profile of various single doses of Fc-Abdeg in cynomolgus monkeys. Figure 20 shows the results of a repeated dose study measuring the effect of various repeated doses of Fc-Abdeg on serum levels of endogenous IgG levels in cynomolgus monkeys. Figure 21 shows the pharmacokinetic profile of various repeated doses of Fc-Abdeg in cynomolgus monkeys. Figure 22 shows the results of a serial dosing study measuring the effect of various doses of Fc-Abdeg on serum levels of endogenous IgG levels in cynomolgus monkeys. Figure 23 shows 24 hours after intravenous loading of Fc-Abdeg at a dose of 20 mg/kg in cynomolgus monkey subjects, followed by daily subcutaneous administration of 3 mg/kg Fc-Abdeg for 28 days, followed by a 32-day treatment-free period. This shows the results of a continuous administration study to measure the effectiveness of this drug.

(detailed explanation)
The present disclosure provides novel methods of reducing serum levels of Fc-containing agents (eg, antibodies and immunoadhesins) in a subject. These methods generally involve administering to the subject an effective amount of an isolated FcRn antagonist that specifically binds FcRn with increased affinity and reduced pH dependence compared to the native Fc region. including. The disclosed methods are particularly useful for treating antibody-mediated disorders such as autoimmune diseases.

(I. Definition)
Unless otherwise defined herein, scientific and technical terms used in connection with the present invention shall have the meanings that are commonly understood by one of ordinary skill in the art. The meaning and scope of terms should be clear, but in the event of hidden ambiguity, the definitions provided herein will be interpreted in advance of any dictionary or external definitions. Further, unless the context otherwise requires, singular terms shall include pluralities and plural terms shall include the singular. Generally, the terminology used with respect to cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein chemistry and nucleic acid chemistry and hybridization described herein, and the techniques thereof, are within the skill of the art. These are terminology and techniques that are well known and commonly used in the technical field.

In order to more easily understand the present invention, certain terms will first be defined.

As used herein, the term "FcRn antagonist" refers to an Fc region (e.g., variant Fc (provided that the drug is not a full-length IgG antibody)
.

The term "Fc region" as used herein refers to the portion of a native immunoglobulin that is formed by the Fc domains of its two heavy chains. The native Fc region is homodimeric.

The term "variant Fc region" as used herein refers to an Fc region that has one or more changes compared to a native Fc region. Changes include amino acid substitutions, additions and/or deletions;
Mention may be made of linking additional components and/or altering the native glycans. The term encompasses heterodimeric Fc regions in which each of the component Fc domains is different. Examples of such heterodimeric Fc regions include, but are not limited to, the "knobs and holes" described in US 8216805, which is incorporated herein by reference in its entirety.
There is an Fc region that is created using technology. The term also refers to a single chain whose component Fc domains are linked together by a linker moiety, as described, for example, in US20090252729A1 and US20110081345A1, each of which is incorporated herein by reference in its entirety.
It also includes the Fc region.

The term "Fc domain" as used herein refers to the portion of a single immunoglobulin heavy chain that begins in the hinge region just upstream of the papain cleavage site and ends at the C-terminus of the antibody. Thus, a complete Fc domain includes at least a portion of the hinge domain (e.g., top, middle, and
or lower hinge region), a CH2 domain, and a CH3 domain.

The term "FcRn binding fragment" as used herein refers to a portion of an Fc region sufficient to confer FcRn binding.

As used herein, the term "EU location" refers to Edelman, GM et al., Proc. Natl. Acad.
USA, 63, 78-85 (1969) and “Sequences of Proteins Subject to Immunology.”
teins of Immunological Interest)” US Dept. Health and Human Services, 5th edition,
Refers to the amino acid position in the EU numbering convention for Fc regions as described in Kabat et al., 1991.

As used herein, the term "CH1 domain" refers to the first (most amino-terminal) constant region domain of an immunoglobulin heavy chain extending from approximately EU positions 118-215. The CH1 domain is adjacent to the VH domain and the amino terminus to the hinge region of the immunoglobulin heavy chain molecule and does not form part of the immunoglobulin heavy chain Fc region.

The term "hinge region" as used herein refers to the portion of a heavy chain molecule that joins the CH1 domain with the CH2 domain. This hinge region contains approximately 25 residues and is flexible, thus allowing the two N-terminal antigen-binding regions to move independently. The hinge region is divided into three distinct domains: the upper, middle, and lower hinge domains (Roux et al., J. Immunol. 161: 4083 (1998)
) can be subdivided into FcRn antagonists of the present disclosure can include all or part of the hinge region.

The term "CH2 domain" as used herein refers to the portion of a heavy chain immunoglobulin molecule that extends from approximately EU positions 231-340.

As used herein, the term "CH3 domain" includes the portion of a heavy chain immunoglobulin molecule that extends approximately 110 residues from the N-terminus of the CH2 domain, e.g., from approximately positions 341 to 446 (EU numbering system). .

The term "FcRn" as used herein refers to neonatal Fc receptor. An exemplary FcRn molecule is
Contains human FcRn encoded by the FCGRT gene described in RefSeq NM_004107.

The term "CD16" as used herein refers to the FcγRIII Fc receptor required for antibody-dependent cell-mediated cytotoxicity (ADCC). Exemplary CD16 molecules include human CD16a described in RefSeq NM_000569.

The term "free cysteine" as used herein refers to a native or engineered cysteine amino acid residue that is present in a substantially reduced form in a mature FcRn antagonist.

As used herein, the term "antibody" refers to an immunoglobulin molecule containing four polypeptide chains, two heavy (H) chains and two light (L) chains interconnected by disulfide bonds; Refers to multimers (e.g., IgM) of Each heavy chain includes a heavy chain variable region (abbreviated VH) and a heavy chain constant region. The heavy chain constant region has three domains, CH1, CH2 and CH3
including. Each light chain includes a light chain variable region (abbreviated VL) and a light chain constant region. The light chain constant region contains one domain (CL). VH and VL areas are framework areas (FR)
It can be further subdivided into regions of hyperdegeneration, termed complementarity determining regions (CDRs), interspersed with more conserved regions termed CDRs.

As used herein, the term "N-linked glycan" refers to the sequon present in the CH2 domain of the Fc region (i.e., the Asn-X-Ser or Asn-X-Thr sequence, where X is any refers to an N-linked glycan attached to the nitrogen (N) in the side chain of asparagine in the amino acid (amino acid). Such N-glycans are, for example, incorporated herein by reference in their entirety.
References Drickamer K and Taylor ME (2006) Introduction to Glycobiology.
biology), 2nd edition.

As used herein, the term "afucosylated" refers to N-linked molecules lacking a core fucose molecule, such as those described in US8067232, the entire contents of which are incorporated herein by reference. Refers to type glycans.

As used herein, the term "bisecting GlcNac" refers to an N-acetylglucosamine linked to a core mannose molecule, such as described in US8021856, the contents of which are incorporated by reference in their entirety. (GlcNAc) refers to N-linked glycans with molecules.

The term "antibody-mediated disorder" as used herein refers to any disease or disorder caused or exacerbated by the presence of antibodies in a subject.

The term "Fc-containing agent" as used herein is any molecule that includes an Fc region.

The term "leukocyte depleting agent" as used herein refers to an agent that upon administration lowers the number of white blood cells in a subject.

The term "B cell depleting agent" as used herein refers to an agent that upon administration reduces the number of B cells in a subject.

The term "T cell depleting agent" as used herein refers to an agent that upon administration reduces the number of T cells in a subject.

The term "autoimmune channelopathies" as used herein refers to diseases caused by autoantibodies against subunits of ion channels or molecules that modulate the channels.

The terms "treat,""treating," and "treatment" as used herein refer to therapeutic or prophylactic measures as described herein. A method of "treatment" includes treating a subject, e.g., a subject with an IL-6-associated disease or disorder (e.g., inflammation and cancer), or a subject predisposed to having such a disease or disorder, with treatment of the disease or disorder, or recurrence. to prevent, cure, delay, reduce the severity of, or ameliorate one or more symptoms of a disease or disorder that has been treated, or to prolong the survival of a subject beyond that expected without such treatment. For this purpose, administration of the antibody of the present invention or antigen-binding fragment thereof is utilized. The term "subject" as used herein includes any human or non-human animal.

As used herein, the term "immunoadhesin" refers to a binding protein (
Refers to an antibody-like molecule that contains a functional domain (eg, a receptor, a ligand, or a cell adhesion molecule).

(II. Method for reducing serum levels of Fc-containing agents)
In one aspect, the present disclosure provides a method of reducing serum levels of Fc-containing agents (e.g., antibodies and immunoadhesins) in a subject, the method comprising: The method comprises administering an effective amount of an isolated FcRn antagonist (e.g., an FcRn antagonist disclosed herein) that specifically binds FcRn with reduced pH dependence.

As shown herein, administration of FcRn antagonists to the subject at doses of about 0.2 to about 200 mg/kg is unexpectedly effective. Thus, in certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 0.2 to about 200 mg/kg (eg, 0.2-200 mg/kg). In certain embodiments, the FcRn antagonist is about 0.2, 2, 20, 70, or 200 mg
/kg (eg, 0.2, 2, 20, 70, or 200 mg/kg). In certain embodiments, the FcRn antagonist is administered to the subject at a dose of about 20 mg/kg (eg, 20 mg/kg).

As shown herein, multiple repeated dosing regimes are unexpectedly superior to single dosing. Thus, in certain embodiments, the FcRn antagonist is administered to the subject at least twice every 20 days. In certain embodiments, the FcRn antagonist is 1, 2, 3
, once every 4, 5, 6, 7, 8, 9, or 10 days. In certain embodiments, the FcRn antagonist is administered at 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 on day 20.
, 16, 17, 18, 19, or 20 times to the subject. In certain embodiments, the FcRn antagonist is administered to the subject once every four days. In certain embodiments, the FcRn
antagonist every 4 days for 13 days (i.e. on days 1, 5, 9, and 13)
administered to the subject. In certain embodiments, 20 mg/kg of FcRn antagonist is administered to the subject every 4 days for 13 days (ie, days 1, 5, 9, and 13).

The FcRn antagonist can be administered to the subject by any means. Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition can be administered, for example, by infusion or bolus injection. In certain embodiments, the FcRn antagonist is administered by intravenous infusion.

The methods disclosed herein can reduce serum levels of any Fc-containing agent. In certain embodiments, the Fc-containing agent is an antibody or an immunoadhesin. In certain embodiments, the Fc-containing agent is a therapeutic or diagnostic agent. In certain embodiments,
The Fc-containing agent is a contrast agent. In certain embodiments, the Fc-containing agent is an antibody drug conjugate. In certain embodiments, the Fc-containing agent is a pathogenic antibody, such as an autoantibody. In certain embodiments, the subject has an antibody-mediated disease or disorder. In certain embodiments, the antibody-mediated disease or disorder is associated with autoantibodies.

Reducing serum levels of Fc-containing agents (eg, antibodies and immunoadhesins) has particular applicability in the treatment of antibody-mediated disorders (eg, autoimmune diseases). Accordingly, in one aspect, the present disclosure provides a method of treating a subject with an antibody-mediated disorder (e.g., an autoimmune disease), comprising administering to the subject an effective amount of an FcRn antagonist composition disclosed herein. The method comprises administering.

Any antibody-mediated disorder can be treated using the methods disclosed herein. In certain embodiments, the antibody-mediated disorder is a disorder amenable to treatment with IVIG. In certain embodiments, the antibody-mediated disorder is an autoimmune disease. Non-limiting autoimmune diseases include allogeneic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune Addison's disease, Alzheimer's disease, anti-neutrophil cytoplasmic autologous Antibodies (ANCA)
, autoimmune diseases of the adrenal glands, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis,
Autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman syndrome, celiac disease - dermatitis, chronic fatigue immunodeficiency syndrome, chronic inflammatory demyelinating polyneuritis (CIDP),
Churg-Strauss syndrome, cicatricial pemphigoid, Crest syndrome, cold agglutinin disease, Crohn's disease, dermatomyositis, discoid lupus erythematosus, essential mixed cryoglobulinemia, stage VIII
factor deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre, Goodpasture syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic pulmonary fibrosis , idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, IgM polyneuropathy, immune-mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen planus, lupus erythematosus, Meniere's disease, mixed connective tissue disease, multifocal sclerosis, type 1 diabetes, multifocal motor neuropathy (M
MN), myasthenia gravis, paraneoplastic bullous pemphigoid, pemphigus vulgaris, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymuscularis rheumatica Pain syndrome, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, solid Organ transplant rejection, stiff man syndrome, systemic lupus erythematosus, Takayasu arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (SJ)
S), temporal arteritis/giant cell arteritis, thrombotic thrombocytopenic purpura, ulcerative colitis, uveitis, dermatitis herpeticum vasculitis, anti-neutrophil cytoplasmic antibody-associated vasculitis, I have vitiligo and Wegener's granulomatosis.

In certain embodiments, the autoimmune disease is an autoimmune channelopathy. Non-limiting channelopathies include neuromyelitis optica, Lambert-Eaton myasthenia syndrome, myasthenia gravis, anti-N-methyl-D-aspartate (NMDA) receptor encephalitis, anti-α-amino-3-hydroxy -5-Methyl-4-isoxazolepropionic acid (AMPA) receptor encephalitis, Morvan syndrome, and glycine receptor antibody-related disorders.

The disclosed methods are particularly suitable for treating antibody-mediated disorders characterized by overproduction of serum immunoglobulins. Accordingly, in certain embodiments, the FcRn antagonist compositions are used to treat hypergammaglobulinemia.

Additionally, the disclosed methods can be used in combination with one or more additional therapeutic agents.
In certain embodiments, the additional therapeutic agent is an anti-inflammatory agent. Any inflammatory agent can be used in combination with the compositions disclosed herein. In certain embodiments, the therapeutic agent is rituximab, daclizumab, basiliximab, muronomab-cd3, infliximab, adalimumab, omalizumab, efalizumab, natalizumab, tocilizumab,
eculizumab, golimumab, canakinumab, ustekinumab, or belimumab. In certain embodiments, the additional therapeutic agent is a leukocyte depleting agent (eg, a B cell or T cell depleting agent). Any leukapheresis agent can be used in combination with the FcRn antagonist compositions disclosed herein. In certain embodiments, the leukocyte depleting agent is a B cell depleting agent. In certain embodiments, the leukapheresis agent is an antibody against a cell surface marker. Suitable cell surface markers include, but are not limited to, CD10, CD19, CD20, CD21.
, CD22, CD23, CD24, CD37, CD53, CD70, CD72, CD74, CD75, CD77, CD79a, CD79b, CD80
, CD81, CD82, CD83, CD84, CD85, or CD86. The FcRn antagonist and the additional therapeutic agent(s) can be administered to the subject via the same or different route(s) of administration, simultaneously or sequentially.

The disclosed methods are also well suited for rapidly reducing serum levels of Fc-containing agents in a subject. Such rapid clearance is advantageous when the Fc-containing agent (eg, an antibody-drug conjugate or an agent that is immunogenic) is toxic because it reduces the subject's exposure to the drug. Rapid clearance is also advantageous when the Fc-containing agent is a contrast agent that requires the agent to be at low serum levels to facilitate imaging. Accordingly, in certain embodiments, the FcRn antagonist composition is used to reduce serum levels of an Fc-containing agent (eg, a contrast agent) in a subject administered the Fc-containing agent. Serum levels of any Fc-containing agent (e.g., therapeutic or diagnostic agent) can be determined by determining the FcRn
can be reduced using antagonist compositions. Non-limiting examples of Fc-containing agents include:
Contrast agents (eg, labeled antibodies), antibody-drug conjugates, or immunogenic agents (eg, non-human antibodies or immunoadhesins). The FcRn antagonist can be administered simultaneously with the Fc-containing agent or sequentially (eg, before or after the Fc-containing agent).

Furthermore, in diseases or conditions that require the administration of a therapeutic agent, the subject often produces antibodies (e.g., anti-drug antibodies) against the therapeutic agent, which in turn can be utilized for the therapeutic purpose for which the therapeutic agent was intended. or cause an adverse reaction in the subject. Accordingly, the methods disclosed herein provide antibodies against a therapeutic agent produced within a subject's body (
For example, it can also be used to remove anti-drug antibodies).

The methods disclosed herein can also be used in combination with therapeutic proteins to enhance the benefits of therapeutic proteins by reducing the levels of IgG; Contributes to decreasing protein bioavailability. In certain embodiments, the present disclosure provides a method of treating a subject with a disorder caused by an immune response to a blood coagulation factor, the method comprising administering to the subject a therapeutically effective amount of an FcRn antagonist composition disclosed herein. The method includes: Suitable blood clotting factors include, but are not limited to, fibrinogen, prothrombin, factor V, factor VII, factor VIII, factor IX, factor X, factor XI, factor XII, factor XIII, or There is the von Willebrand factor. This method can be used, for example, to modulate or treat or prevent immune responses to blood clotting factors in patients suffering from hemophilia A or hemophilia B. In certain embodiments, the method can be used, for example, to modulate or treat the immune response to therapeutic erythropoietin in a patient suffering from aplasia erythropoietin (PRCA).

FcRn contributes to the transport of maternal antibodies across the placenta to the fetus in pregnant women. Therefore,
When a pregnant woman is given an Fc-containing agent (e.g., a therapeutic antibody), the agent may transfer Fc across the placenta.
There is a risk of contact with the fetus as a result of Rn-mediated transport. In order to avoid any potential adverse effects of the Fc-containing agent on fetal development, it would be beneficial to block the function of FcRn. Accordingly, the present disclosure provides a method of preventing placental transfer of an Fc-containing agent (e.g., a therapeutic antibody) to a fetus in a pregnant woman, the method comprising administering to said woman an FcRn antagonist composition disclosed herein. The method comprises administering the agent simultaneously or sequentially (before or after) with the agent.

The methods disclosed herein are also applicable to inflammatory disorders including, but not limited to, asthma, ulcerative colitis and inflammatory bowel syndrome, allergic rhinitis/sinusitis, skin allergies (
Allergies including urticaria/hives, angioedema, atopic dermatitis), food allergies, drug allergies, insect allergies, mastocytosis, osteoarthritis, rheumatoid arthritis, and spondyloarthropathies. Can be used to treat arthritis including.

The successful implementation of gene therapy for the treatment of a disease or condition can be hampered by the production of antibodies specific for the therapeutic protein encoded by the transgene and possibly the vector used to deliver the transgene. . Accordingly, when the FcRn antagonist compositions disclosed herein are administered in combination with gene therapy, the benefits of the encoded therapeutic protein can be enhanced by reducing the levels of IgG. These methods are particularly useful in situations where IgG antibodies contribute to reduced bioavailability of gene therapy vectors or encoded therapeutic proteins. The gene therapy vector is, for example,
It can be a viral vector such as adenovirus and adeno-associated virus.
Diseases that can be treated using gene therapy include, but are not limited to, cystic fibrosis, hemophilia, PRCA, muscular dystrophy, or lysosomal storage diseases such as Gaucher disease and Fabry disease.

Any subject can be treated using the methods disclosed herein. In certain embodiments, the subject is a human or a cynomolgus monkey.

(III. FcRn antagonist)
The methods disclosed herein generally include administering to a subject an effective amount of an isolated FcRn antagonist, wherein the FcRn antagonist has increased and decreased affinity as compared to the native Fc region. It specifically binds to FcRn with a certain pH dependence. Generally, these
FcRn antagonists include variant Fc regions or FcRn-binding fragments thereof that specifically bind FcRn with increased affinity and reduced pH dependence as compared to the native Fc region.
The FcRn antagonist can be combined with Fc-containing agents (e.g., antibodies and immunoadhesins) in vivo.
Inhibits binding of Rn, thereby increasing the rate of degradation of Fc-containing agents and concomitantly reducing serum levels of these agents.

As shown herein, certain isolated variant Fc regions (e.g., EU position 252 and
, 254, 256, 433, 434, and 436) are more effective FcRn antagonists in vivo than full-length antibodies containing the same variant Fc regions. be. Therefore, in certain embodiments, the FcRn antagonist composition is not a full-length antibody. In certain embodiments, the FcRn antagonist composition does not include antibody variable domains. In certain embodiments, the FcRn antagonist composition does not include an antibody variable domain or a CH1 domain. However, in certain embodiments, the FcRn antagonist composition can include a variant Fc region linked to one or more additional binding domains or binding moieties, including antibody variable domains.

Any Fc region can be altered to produce variant Fc regions for use in the FcRn antagonist compositions disclosed herein. Generally, the Fc region or its FcRn
The binding fragment is derived from human immunoglobulin. However, the Fc region may be used in, for example, camelid species, rodents (e.g. mice, rats, rabbits, guinea pigs) or non-human primates (
It will be appreciated that immunoglobulins may be derived from any other mammalian species, including (eg, chimpanzee, macaque) species. Furthermore, the Fc region or portion thereof may be IgM, IgG, IgD, IgA
and any immunoglobulin class, including IgE, and any immunoglobulin isotype, including IgG1, IgG2, IgG3 and IgG4. In certain embodiments, the Fc region is an IgG Fc region (eg, a human IgG region). In certain embodiments, the Fc region is an IgG
1 Fc region (eg, human IgG1 region). In certain embodiments, the Fc region is a chimeric Fc region that includes portions of several different Fc regions. Suitable examples of chimeric Fc regions are described in US20110243966A1, which is incorporated herein by reference in its entirety. various F
c region gene sequences (eg, human constant region gene sequences) are available in public deposit format. It will be understood that the scope of the invention includes alleles, variants and mutations of the Fc region.

The Fc region can be further truncated or deleted internally to produce its minimal FcRn binding fragment. The ability of an Fc region fragment to bind FcRn can be determined using any art-recognized binding assay, such as ELISA.

To enhance the manufacturability of the FcRn antagonists disclosed herein, the components
Preferably, the Fc region does not contain any cysteine residues that do not form disulfide bonds. Thus, in certain embodiments, the Fc region does not contain free cysteine residues.

FcRn with increased affinity and reduced pH dependence compared to the native Fc region.
Any Fc variant or FcRn-binding fragment thereof that specifically binds to
Can be used in FcRn antagonist compositions. In certain embodiments, variant Fc regions contain amino acid changes, substitutions, insertions and/or deletions that confer desired properties. In certain embodiments, the variant Fc region or fragment is at EU position 252, 254, 256, 433, respectively.
, 434, and 436 contain amino acids Y, T, E, K, F, and Y. Non-limiting examples of amino acid sequences that can be used in variant Fc regions are provided in Table 1 herein. In certain embodiments, the amino acid sequence of the Fc domain of the variant Fc region comprises the amino acid sequence set forth in SEQ ID NO:1. In certain embodiments, the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3. In certain embodiments, the FcRn antagonist consists of a variant Fc region, wherein the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3.

table 1. Amino acid sequences that are non-limiting examples of variant Fc regions

In certain embodiments, the variant Fc region has been altered for additional Fc receptors (
e.g., increased or decreased binding affinity. The variant Fc region has one or more Fc regions.
γ receptors, such as FcγRI (CD64), FcγRIIA (CD32), FcγRIIB (CD32), FcγRIIIA (CD
16a), and can have an altered (eg, increased or decreased) binding affinity for FcγRIIIB (CD16b). Any art-recognized method of altering affinity for additional Fc receptors can be utilized. In certain embodiments, variant Fc
The amino acid sequence of the region is changed.

In certain embodiments, the variant Fc region is 234, 235, 236, 239, 240, 241, 243, 244, 245, 247, 252, 254, numbered according to the EU index as described in the Kabat reference. twenty five
6, 262, 263, 264, 265, 266, 267, 269, 296, 297, 298, 299, 313, 325, 326, 327, 32
Contains a non-naturally occurring amino acid residue at one or more positions selected from the group consisting of 8, 329, 330, 332, 333, and 334. Optionally, the Fc region can include non-naturally occurring amino acid residues at additional and/or alternative positions known to those skilled in the art (e.g. U.S. Patent No. 5,624,821; No. 6,277,375; No. 6,737,056
PCT Patent Publication No. WO 01/58957; No. WO 02/06919; No. WO 04/016750; No. WO 04/02920
7; see WO 04/035752 and WO 05/040217).

In certain embodiments, the variant Fc region is 234D, 234E, 234N, 234Q, 234T, 234H, 234Y, 234I, 234V, 234F, 23 numbered according to the EU index as described in the Kabat reference.
5A, 235D, 235R, 235W, 235P, 235S, 235N, 235Q, 235T, 235H, 235Y, 235I, 235V, 235F
, 236E, 239D, 239E, 239N, 239Q, 239F, 239T, 239H, 239Y, 2401, 240A, 240T, 240M,
241W, 241 L, 241Y, 241E, 241R, 243W, 243L 243Y, 243R, 243Q, 244H, 245A, 247V, 24
7G, 252Y, 254T, 256E, 262I, 262A, 262T, 262E, 263I, 263A, 263T, 263M, 264L, 264I
, 264W, 264T, 264R, 264F, 264M, 264Y, 264E, 265G, 265N, 265Q, 265Y, 265F, 265V,
265I, 265L, 265H, 265T, 266I, 266A, 266T, 266M, 267Q, 267L, 269H, 269Y, 269F, 26
9R, 296E, 296Q, 296D, 296N, 296S, 296T, 296L, 296I, 296H, 269G, 297S, 297D, 297E
, 298H, 298I, 298T, 298F, 299I, 299L, 299A, 299S, 299V, 299H, 299F, 299E, 313F,
325Q, 325L, 325I, 325D, 325E, 325A, 325T, 325V, 325H, 327G, 327W, 327N, 327L, 32
8S, 328M, 328D, 328E, 328N, 328Q, 328F, 328I, 328V, 328T, 328H, 328A, 329F, 329H
, 329Q, 330K, 330G, 330T, 330C, 330L, 330Y, 330V, 330I, 330F, 330R, 330H, 332D,
Contains at least one non-naturally occurring amino acid residue selected from the group consisting of 332S, 332W, 332F, 332E, 332N, 332Q, 332T, 332H, 332Y, and 332A. Optionally, the Fc region can include additional and/or alternative non-naturally occurring amino acid residues known to those skilled in the art (
For example, U.S. Patent No. 5,624,821, the entire contents of which are incorporated herein by reference;
No. 6,277,375; No. 6,737,056; PCT Patent Publication No. WO 01/58957; No. WO 02/06919; No. WO
04/016750; WO 04/029207; WO 04/035752 and WO 05/040217).

Other known Fc variants that can be used in the FcRn antagonists disclosed herein include, without limitation, Ghet, the entire contents of which are incorporated herein by reference.
ie et al., 1997, Nat. Biotech. 15:637-40; Duncan et al., 1988, Nature 332:563-
564; Lund et al., 1991, J. Immunol., 147:2657-2662; Lund et al., 1992, Mol. Im
munol., 29:53-59; Alegre et al., 1994, Transplantation 57:1537-1543; Hutchins et al., 1995, Proc Natl. Acad Sci USA, 92:11980-11984; Jefferis et al., 1995, I
mmunol Lett., 44:111-117; Lund et al., 1995, Faseb J., 9:115-119; Jefferis et al., 1996, Immunol Lett., 54:101-104; Lund et al., 1996 , J. Immunol., 157:4963
-4969; Armour et al., 1999, Eur J Immunol 29:2613-2624; Idusogie et al., 2000,
J. Immunol., 164:4178-4184; Reddy et al., 2000, J. Immunol., 164:1925-1933; Xu
et al., 2000, Cell Immunol., 200:16-26; Idusogie et al., 2001, J. Immunol.,
166:2571-2575; Shields et al., 2001, J Biol. Chem., 276:6591-6604; Jefferis et al., 2002, Immunol Lett., 82:57-65; Presta et al., 2002, Biochem Soc Trans., 3
0:487-490); U.S. Patent No. 5,624,821; No. 5,885,573; No. 5,677,425; No. 6,165,745;
No. 6,277,375; No. 5,869,046; No. 6,121,022; No. 5,624,821; No. 5,648,260; No. 6,5
28,624; 6,194,551; 6,737,056; 6,821,505; 6,277,375; U.S. Patent Publication No. 2004/0002587 and PCT Publication No. WO 94/29351; WO 99/58572; ;
WO 02/060919; WO 04/029207; WO 04/099249; WO 04/063351.

In certain embodiments, the variant Fc region is a heterodimer, where the component Fc domains are different from each other. Methods for producing Fc heterodimers are known in the art (e.g., US 8216805, herein incorporated by reference in its entirety).
Please refer to ). In certain embodiments, the variant Fc region is a single chain Fc region, where the component Fc domains are joined together by a linker moiety. Methods for producing single chain Fc regions are known in the art (see, eg, US20090252729A1 and US20110081345A1, each of which is incorporated by reference in its entirety).

Pathogenic IgG antibodies observed in autoimmune diseases may trigger the pathogenesis of these diseases or contribute to disease progression and promote disease via inappropriate activation of cellular Fc receptors. It is thought that it will become a medium for Aggregated autoantibodies and/or autoantibodies complexed with autoantigens (immune complexes) bind to activated Fc receptors and cause numerous autoimmune diseases (these are partially (e.g., Clarkson et al., NEJM 314(9), 1236-1239 (2013), each incorporated herein by reference in its entirety); US20040010124A1; US20040047862A1; and US2004
/0265321A1). Therefore, to treat antibody-mediated disorders (e.g., autoimmune diseases), it is important to remove harmful autoantibodies and to activate these antibodies' activated Fc receptors (e.g., Fcγ such as CD16a). Both would be beneficial.

Accordingly, in certain embodiments, the variant Fc region of the FcRn antagonist is CD16a
(e.g., human CD16a). This is particularly advantageous in allowing the FcRn antagonist to further antagonize the immune complex-induced inflammatory response of autoantibodies that are targeted for removal by inhibition of FcRn. CD16a (e.g. human CD16a
) can be utilized.
In certain embodiments, the FcRn antagonist comprises a variant Fc region that includes an N-linked glycan (eg, at EU position 297). In this case, changing the glycan structure may increase the binding affinity of the FcRn antagonist for CD16a. F
N-linked glycan changes in the c region are well known in the art. For example, afucosylated N-
Linked glycans or N-glycans with bisecting GlcNac structures have been shown to exhibit increased affinity for CD16a. Thus, in certain embodiments, the N-linked glycans are afucosylated. Afucosylation can be accomplished using any art-recognized method. For example, an FcRn antagonist can be expressed in cells deficient in fucosyltransferase such that fucose is not added to the N-linked glycan at EU position 297 of the variant Fc region (e.g., its entire content is authentic). (See US 8,067,232, incorporated by reference). In certain embodiments, the N-linked glycan has a bisecting GlcNac structure. The bisecting GlcNac structure can be achieved using any art-recognized method. For example, FcRn antagonists are β1-4-N-acetylglucosaminyltransferase III (G
nTIII), such that a bisecting GlcNac is added to the N-linked glycan at EU position 297 of the variant Fc region (e.g., the entire contents of which are herein (see US 8021856, incorporated by reference). moreover,
Alternatively, changes in the N-linked glycan structure can also be achieved by in vitro enzymatic methods.

In certain embodiments, the FcRn antagonist comprises a plurality of FcRn antagonist molecules, wherein at least 50% (optionally, at least 60,
70, 80, 90, 95, or 99%) contains a variant Fc region containing an afucosylated N-linked glycan at EU position 297, or an FcRn-binding fragment thereof.

In certain embodiments, the FcRn antagonist comprises a plurality of FcRn antagonist molecules, wherein at least 50% (optionally, at least 60,
70, 80, 90, 95, or 99%) contain a variant Fc region containing an N-linked glycan with a bisecting GlcNac at EU position 297, or an FcRn-binding fragment thereof.

In certain embodiments, the variant Fc region does not include N-linked glycans. This can be accomplished using any art-recognized method. For example, the Fc variant can be expressed in cells that are incapable of N-linked glycosylation. Additionally or alternatively, the amino acid sequence of the Fc variant can be altered (eg, by mutation of the NXT sequon) to prevent or inhibit N-linked glycosylation. Or,
Fc variants can be synthesized in cell-free systems (eg, chemical synthesis).

In certain embodiments, FcRn can be isolated, e.g., by covalently attaching a molecule (e.g., a binding or imaging moiety) to an FcRn antagonist such that the covalent bond does not prevent the FcRn antagonist from specifically binding to FcRn. Antagonist molecules can be modified. For example, and without limitation, the FcRn antagonist may include glycosylation, acetylation,
Modifications can be made by pegylation, phosphorylation, amidation, derivatization with known protective blocking groups, proteolytic cleavage, conjugation with cellular ligands or other proteins, and the like.

In certain embodiments, the FcRn antagonist comprises a variant Fc region linked to a half-life extender. The term "half-life extender" as used herein refers to any molecule that, when linked to an FcRn antagonist disclosed herein, increases the half-life of the FcRn antagonist. Any half-life extender can be linked (covalently or non-covalently) to the FcRn antagonist. In certain embodiments, the half-life extender is polyethylene glycol or human serum albumin. In certain embodiments, the FcRn antagonist is a binding molecule that specifically binds to a half-life extender present in a subject, such as serum albumin (e.g., human serum albumin).
, IgG, red blood cells, etc., are linked to molecules or cells carried by the blood.

(IV. Pharmaceutical composition)
In certain embodiments, the method comprises an FcRn antagonist or an FcRn antagonist disclosed herein.
A pharmaceutical composition comprising an Rn antagonist composition and a pharmaceutically acceptable carrier or excipient is utilized. Examples of suitable pharmaceutical carriers are listed in Remington's Pharmaceutical Science.
ces)” (ed. E. W. Martin). Examples of excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dry skim milk, glycerol, propylene glycol, Examples include water and ethanol. The composition can also include a pH buffering agent, and a wetting or emulsifying agent.

The pharmaceutical composition can be formulated for parenteral administration (eg, intravenous or intramuscular) by bolus injection. Formulations for injection may be presented in unit dosage form, eg, in ampoules or in multi-dose containers, with an added preservative. The compositions can take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles;
Formulatory agents such as stabilizing and/or dispersing agents may be included. Alternatively, the active ingredient can be in powder form for constitution with a suitable vehicle, eg pyrogen-free water.

In the disclosed methods, FcRn antagonists can be linked to chelating agents, such as those described in US Pat. No. 5,326,856. The peptide-chelator conjugate can be subsequently radiolabeled to provide a contrast agent for the diagnosis or treatment of diseases or conditions involving modulation of IgG levels.

(V. Examples)
The invention is further illustrated by the following examples, which should not be construed as further limiting. The contents of the sequence listing, drawings, and all cited references, patents, and published patent applications cited throughout this application are expressly incorporated herein by reference.

(Example 1: Effect of Fc-Abdeg on serum IgG levels in cynomolgus monkeys)
Human anti-lysozyme IgG (HEL-Abdeg) on serum IgG levels of tracer antibodies and human containing amino acids Y, T, E, K, F, and Y at EU positions 252, 254, 256, 433, 434, and 436, respectively.
The effects of the IgG Fc region (Fc-Abdeg; SEQ ID NO: 2) were determined in cynomolgus monkeys. Specifically, cynomolgus monkeys were given 1 mg/kg of anti-mouse CD70 hIgG1 tracer antibody (FR70-hI
gG1; Oshima et al., Int Immunol 10(4): 517-26 (1998)) was administered by intravenous bolus injection. Five minutes later, animals were infused with either 7 mg/kg Fc-Abdeg, 20 mg/kg HEL-Abdeg, or PBS (2 monkeys per group). The infusion was carried out within 1 hour and the animals received a volume of 10 ml/kg. Blood samples (3 x 150 μl) were taken 5 minutes before dosing (“pre-dose”) and at 5 minutes, 2 hours, 6 hours, 24 hours, 48 hours, 72 hours, 96 hours and 120 hours after the end of the infusion. It was collected after an hour.
Tracer levels were determined by performing mCD70 binding ELISA and data were plotted against tracer levels at the end of dosing (Figure 1). We also determined total cynomolgus IgG levels (Figure 2). These experimental results indicate that Fc-Abdeg reduced tracer antibodies more efficiently than an equivalent amount of HEL-Abdeg.

In addition to its important role in the IgG recycling pathway, FcRn is also involved in albumin homeostasis (Chaudhury et al., J Exp Med. 197(3):315-22 (2003)). FcRn interacts with IgG-Fc and albumin at different sites, and binding can occur simultaneously (Andersen et al., Nat Co.
mmun. 3:610 (2012)). Conceptually, by blocking IgG recycling using Abdeg-engineered molecules, albumin-FcRn interactions should not be interfered with. This hypothesis was confirmed in in vivo studies in mice. Here, the authors showed that hIgG1 molecules with Abdeg have no effect on albumin levels (Patel et al., J Immunol 187(2): 1015-22 (20
11)). In the experiments described above, albumin levels were also determined at -3, 3 and 17 days after the end of the infusion. Similar to the study in mice, no significant changes in albumin levels were observed after Fc-Abdeg or HEL-Abdeg treatment (see Figure 3).

In subsequent experiments, the antibody removal efficacy of Fc-Abdeg was compared to IVIG. Specifically, 70 mg/kg
Cynomolgus monkeys were administered 1 mg/kg tracer antibody (FR70-hIgG1) 2 days before dosing with Fc-Abdeg or 2 g/kg IVIG (2 monkeys per group). Infusions of Fc-Abdeg and IVIG were performed within 4 hours and animals were administered a volume of 20 ml/kg. Blood samples (3 × 150 μl) were collected 5 minutes before dosing (“pre-dose”) and at 5 minutes, 2 hours, 6 hours, 24 hours, 48 hours, 72 hours, and 96 hours after the end of the infusion.
Samples were taken after 120 hours, 120 hours, and 168 hours. Tracer levels were determined by mCD70 binding ELISA and plotted against pre-dose levels (Figure 4). Compared to the clinical dose (2 g/kg) of IVIG treatment, 70 mg/kg Fc-Abdeg showed significantly enhanced tracer clearance kinetics and could also be eliminated more efficiently (Abdeg , >95% tracer clearance in 4 days vs. ~75% clearance in 7 days for IVIG).

(Example 2: Effect of afucosylation on the affinity of Fc-Abdeg for human CD16a and mouse CD16-2)
The binding affinity of Fc-Abdeg to hCD16a was determined and compared to the afucosylated form (Fc-Abdeg-POT). The same experiment included Fc-Abdeg variants (“Fc-
Abdeg-S239D/I332E”). Specifically, we prepared Maxisorp plates with 100 ng/well of Neut
It was coated with ravidin biotin binding protein (ThermoScientific, 31000) and incubated overnight at 4°C. The next day, the plates were blocked with PBS+1% casein for 2 hours at room temperature. Next, biotin-labeled hCD16a (Sino Biological, 10389-H27H1-B)
Add 100 μl/well of 0 ng/ml solution (diluted in PBS + 0.1% casein) to the plate and
A concentration gradient (1 μM to 0.005 nM) of deg or Fc-Abdeg-POT molecules was incubated for 1 hour at room temperature before application and an additional 1 hour after application. Binding to hCD16a was determined using HRP-conjugated polyclonal goat anti-human Fc antibody (Jackson ImmunoResearch, 109-035-008) (incubated for 1 hour at room temperature, diluted 1/50,000 in PBS + 0.1% casein), followed by
Detection was performed by adding μl of TMB equilibrated to room temperature (SDT reagent #s TMB). Plates were incubated for 10 minutes, then 100 μl 0.5 NH ₂ SO ₄ was added and OD450nm measurements were taken. EC50 values were determined using GraphPad Prism software. The results of these experiments, described in Figure 5, show that the affinity for hCD16a increases >30-fold when Fc-Abdeg molecules are defucosylated ( _EC50 = for Fc-Abdeg-POT). 13 nM vs. EC ₅₀ >0.4 μM for fucosylated Fc-Abdeg). As expected, the binding affinity of the Fc-Abdeg-S239D/I332E variant to hCD16a was higher compared to wild-type Fc-Abdeg (EC ₅₀ =6 nM).

Using experimental procedures similar to those previously described, mouse CD16-2 (Sino Biological, 50036-
The binding affinity for M27H-B) was determined. The results of these experiments, described in Figure 6, once again demonstrate the higher affinity of the afucosylated variants compared to wild-type Fc-Abdeg.
( _EC50 = 11 nM vs. _EC50 >100 nM). Fc-Abdeg-POT variant mCD16 versus wild-type Fc-Abdeg
The fold increase in affinity for -2 is low compared to the fold increase observed for binding to human CD16a. This effect was not observed for the Fc-Abdeg-S239D/I332E variant (EC ₅₀ = 2 nM), which showed similar affinity for both human and mouse CD16 compared to wild-type Fc-Abdeg. with a fold increase ( _EC50 = 2 nM).

Autoantibodies that have formed a complex with self-antigens bind to activated FcγR, thereby triggering autoimmune diseases. Autoimmune diseases arise in part due to immunologically mediated inflammation of self tissues. The ability of Fc-Abdeg to antagonize autoimmune antibody and FcγRIII receptor interactions on NK cells was evaluated in two ADCC-based assays.

First, using ADCC reporter bioassay (Promega, G7016), Fc-Abdeg, Fc-Abd
The competitive hCD16a binding efficacy of eg-POT and Fc-Abdeg-S239D/I332E was analyzed. Specifically, 10
10,000 CD20-expressing Raji cells (
Target cells) were incubated with 60,000 hCD16a-expressing Jurkat cells (effector cells). Cells were incubated at 37°C for 6 hours, after which the bioluminescent signal, a measure of ADCC activity, was measured. Luciferase signal at 100 ng/m in the absence of competitors
l plotted against the signal obtained with anti-CD20 (see Figure 7). These experiments showed that both Fc-Abdeg-POT and Fc-Abdeg-S239D/I332E efficiently blocked anti-CD20-induced ADCC signals, whereas incubation with wild-type Fc-Abdeg expressed on Jurkat cells. It is shown that there is no competitive binding to hCD16a.

In the following ADCC assay, anti-hCD70 antibody (27B3-hIgG1) by Fc-Abdeg and Fc-Abdeg-POT
The inhibition of the lytic activity of was tested as a criterion for competitive hCD16 binding. Specifically, approximately 50,000 hCD70-expressing U266 cells were treated with 50 ng/ml anti-hCD70 antibody and Fc-Abdeg, Fc-Abdeg-POT and IVIG.
into approximately 300,000 PBMCs that had just been purified from a healthy donor in the presence of a concentration gradient. U266 cells were incubated for 2 days and subsequent cell lysis was performed using a marker specific for U266 cells (CD2).
Analyzed by FACS using 8). The results of these experiments described in Figure 8 demonstrate that anti-CD70 antibodies efficiently lyse U266 cells and that addition of Fc-Abdeg-POT can attenuate this clearance in a dose-dependent manner. We show that this removal cannot be attenuated by either wild-type Fc-Abdeg or IVIG. These data indicate that the Fc-Abdeg POT is similar to wild type Fc-Abdeg and IV.
It has been shown to have enhanced competitive CD16a binding properties compared to IG.

(Example 3: Mouse acute ITP model)
The therapeutic efficacy of Fc-Abdeg, Fc-Abdeg-POT, Fc-Abdeg-S239D/I332E molecules was tested in a mouse model of acute immune thrombocytopenia. Specifically, C57BL/6 mice were injected with I.
VIG (20 mg/animal), Fc-Abdeg (1 mg/animal), Fc-Abdeg-POT (1 mg/animal), Fc-Abdeg-S239D
/I332E (1 mg/animal) or saline (5 animals/group). Prior to treatment, blood samples were taken for baseline platelet count measurements. One hour later, mice were treated with 5 μg/animal of the anti-mouse platelet antibody MWReg30 (Nieswandt et al., Blood 94:684-93 (1999)). Platelet counts were monitored over 24 hours. Platelet counts were normalized to the initial count for each mouse, and platelet counts were determined using flow cytometry via anti-CD61 staining. figure
The results of these experiments, described in 9, showed that pretreatment with Fc-Abdeg reduced MWReg30-induced thrombocytopenia with similar potency compared to a 7-fold higher molar dose of IVIG, as well as , as seen from the improvement in platelet counts at 180 and 1440 minutes.
Blockade of FcγR by T and Fc-Abdeg-S239D/I332E is shown to have synergistic beneficial effects in this model.

(Example 4: Manufacturability of Fc-Abdeg)
CHO cells (
Evitria, Switzerland). Following transfection, high titers of Fc-Abdeg were detected in the supernatant (200-400 mg/ml). Fc-Abdeg in CHO GS-XCEED cell line (Lonza, Great-Britain)
A similar favorable production profile was seen when expressed from an expression construct stably integrated into Stable transfectants produced an average of 3 g/L in a 10 L stirred tank bioreactor;
Several clones producing up to 6 g/L of Fc-Abdeg were identified.

The manufacturability of Fc-Abdeg was further investigated by analysis of aggregates and degradation products after protein A purification from the Fc-Abdeg production runs described above. Specifically, 137 μg of Fc-Abdeg
was applied onto a Superdex 200 10/300 GL gel filtration column (GE Healthcare) connected to an AktaPurifier chromatography system. The results of this experiment, described in Figure 10, showed that only a very small percentage of Fc-Abdeg aggregates (~0.5%) were observed, while no Fc-Abdeg degradation products were detected. Furthermore, applying various stress conditions (freeze-thaw, rotational or temperature stress) to Protein A purified Fc-Abdeg did not result in easily appreciable changes in physicochemical and functional properties. Collectively, these data demonstrate the excellent manufacturability of Fc-Abdeg.

(Example 5: Dose escalation study of Fc-Abdeg in cynomolgus monkeys)
Fc-A in cynomolgus monkeys to determine onset of pharmacokinetic effects as well as saturating doses.
A dose escalation study of bdeg was conducted. For this purpose, 1 mg/kg anti-mouse CD70 hI was administered to cynomolgus monkeys.
gG1 tracer antibody (FR70-hIgG1) was administered by intravenous bolus injection. After 48 hours, animals were infused with various doses of FC-Abdeg (0.2 mg/kg, 2 mg/kg, 20 mg/kg or 200 mg/kg) or vehicle (PBS). The infusion was performed within 3 hours and the animals received a volume of 36.36 ml/kg. Each test group consisted of two animals. 5 minutes before administration (“pre-dose”) and 5 minutes, 2 hours, 6 hours, 24 hours, 48 hours, 72 hours, 5 days, 7 days, 10 days, and 14 days after the end of the infusion. Samples (3 x 150 μl) were taken. Tracer IgG (see Figure 11) and endogenous
Both levels of IgG (see Figure 12) were determined by ELISA and plotted against pre-dose levels. Data from the 70 mg/kg dose was added from the experiment described in Figure 4 herein. Administration of 0.2 mg/kg FC-Abdeg to animals did not significantly affect the rate of tracer clearance and did not affect endogenous IgG levels. A clear pharmacokinetic effect is seen at 2 mg/kg, and this effect levels off at doses above 20 mg/kg. Administration of Fc-Abdeg alone reduces IgG in cynomolgus monkeys by up to 55% within 3 to 4 days.

FcRn binding is restricted to the gamma subtype of immunoglobulins, which may explain the much longer half-life of the gamma subtype compared to other immunoglobulin subtypes. Therefore, blockade of the IgG recycling function of FcRn by Fc-Abdeg should not interfere with the levels of endogenous non-IgG immunoglobulins, a feature that was observed in serum samples of animals treated with 200 mg/kg Fc-Abdeg. (See Figures 13 and 14). Combined with the observations described in Example 1 showing that treatment with Fc-Abdeg does not affect albumin levels, these data demonstrate the specific pharmacokinetic effects of Fc-Abdeg.

Next, the pharmacokinetic profile of Fc-Abdeg was determined by ELISA. As calculated from its PK profile (see Figure 5), Fc-Abdeg has a short half-life (estimated half-life ~1.5 days). At saturating levels, Fc-Abdeg that is not bound to FcRn is efficiently eliminated due to its own mechanism of action. Furthermore, the molecular size of Fc-Abdeg is
Close to the cut-off value for renal clearance (~60 kDa).

(Example 6: Repeated administration test of Fc-Abdeg in cynomolgus monkeys)
A single infusion of Fc-Abdeg at a dose of 20 mg/kg reduces endogenous IgG levels in cynomolgus monkeys by 55% within approximately 3-4 days. Subsequent experiments tested whether repeated administration of Fc-Abdeg would further reduce these levels. Two strategies were used: one group of monkeys received infusions of the test component every 24 hours for the first four days (days 1, 2, 3, and 4), whereas the other group received infusions of the test component every 4 days. (days 1, 5, 9, and 13). For each individual dose, Fc-Abdeg
was administered at a dose of 20 mg/kg. Each test group consisted of two animals. The infusion procedure was the same as that described in Example 1. Clear pharmacodynamic differences between the two different groups are observed at day 7 (see Figure 16). Daily Fc-Abdeg in the first 4 days
administration showed a similar reduction in IgG levels as a single infusion of the same dose (dashed line, overlaid with data from Example 1 as reference). Administration of Fc-Abdeg once every four days results in a greater and longer reduction in these levels. A reduction of up to 25% from initial levels is observed in this treatment group.

The pharmacokinetic profile of Fc-Abdeg was determined (see Figure 7). The profile is consistent with the findings from the dose escalation study described in Example 5.

(Example 7: Further dose escalation study of Fc-Abdeg in cynomolgus monkeys)
To further explore the onset and saturation of pharmacodynamic effects, a follow-up dose escalation study of Fc-Abdeg was conducted in cynomolgus monkeys. For this purpose, animals were given various doses of FC-Abdeg (10 mg/kg, 30
mg/kg, 50 mg/kg and 100 mg/kg) or vehicle (PBS). Infusions were performed within 2 hours and each cohort consisted of 4 animals (2 males and 2 females). Endogenous IgG levels were determined by ELISA and plotted against pre-dose levels (see Figure 18). A moderate pharmacodynamic effect was observed at a dose of 10 mg/kg, and this effect leveled off at doses above 30 mg/kg. The rate of IgG clearance, onset of action, and pharmacokinetic profile (see Figure 19; see Table 2) were comparable to the findings described in Example 5.

＃：Fc-Abdegの血清分析から得られた値、他の全ての値は毒性動態学的分析により算出さ
れた。
－：計算不能又は利用可能なデータから合理的な解釈ができない
DPF：用量比例計数＝[AUC_{0-t ラスト} (x mg/kg)/ AUC_{0-t ラスト} (10 mg/kg)]/ [(x mg/k
g)/ (10 mg/kg)] Table 2: Pharmacokinetic properties of Fc-Abdeg in cynomolgus monkeys

#: Value obtained from serum analysis of Fc-Abdeg, all other values were calculated by toxicokinetic analysis.
-: Incalculable or cannot be rationally interpreted from available data
DPF: Dose proportional count = [AUC _{0-t Last} (x mg/kg)/ AUC _{0-t Last} (10 mg/kg)]/ [(x mg/k
g)/ (10 mg/kg)]

(Example 8: Further repeated administration study of Fc-Abdeg in cynomolgus monkeys)
To further investigate the pharmacodynamic effects of long-term Fc-Abdeg administration, Fc-Abd
A follow-up repeated dose study of eg was conducted. For this purpose, animals were given various concentrations of FC-Abdeg (3 mg/kg
, 30 mg/kg and 100 mg/kg) or vehicle (PBS) were infused every 48 hours for a total of 15 infusions. A 30-day recovery period was included after the treatment period. Each cohort consisted of 4 animals, with the exception of the 100 mg/kg cohort (which consisted of 3 animals).
. Levels of endogenous IgG were determined by ELISA and plotted against pre-dose levels (Figure 20
Please refer to mean ± SEM). A clear decrease in IgG levels was observed at all tested doses;
This pharmacodynamic effect persisted until the end of the treatment period. After the treatment period, IgG levels returned to baseline within 10 days. The pharmacokinetic profile is shown in Figure 21.

(Example 9: Long-term administration test of Fc-Abdeg in cynomolgus monkeys)
Cynomolgus monkeys were infused with an intravenous loading dose of 20 mg/kg FC-Abdeg, followed by daily subcutaneous administration of 1, 3, or 5 mg/kg Fc-Abdeg starting 24 hours after the first dose and continuing for 12 days. was further applied. Each test group consisted of two animals. IgG levels were determined by ELISA and plotted against pre-dose levels (see Figure 22). Data are also presented for one cynomolgus monkey from the 3 mg/kg cohort where treatment lasted longer than 12 days. The monkeys were infused with an intravenous loading dose of 20 mg/kg FC-Abdeg and given daily subcutaneous administration of 3 mg/kg Fc-Abdeg starting 24 hours after the first dose and continuing for 28 days. This was followed by an additional 32-day treatment-free period (see Figure 23). IgG levels decreased by up to 60% of original levels and remained consistently low throughout the treatment period. IgG within 2 weeks after the treatment period
The level returned to baseline level.
The present application provides an invention having the following configuration.
(Configuration 1)
A method of reducing serum levels of an Fc-containing agent in a subject, the method comprising administering to the subject an effective amount of an isolated FcRn antagonist comprising a variant Fc region or an FcRn-binding fragment thereof, the method comprising: Fc domains of EU positions 252, 254, 256, 433, 434, and 436
and wherein the FcRn antagonist is administered to the subject at a dose of about 0.2 to about 200 mg/kg.
(Configuration 2)
A method of reducing serum levels of an Fc-containing agent in a subject, the method comprising administering to the subject an effective amount of an isolated FcRn antagonist comprising a variant Fc region or an FcRn-binding fragment thereof, the method comprising: Fc domains of EU positions 252, 254, 256, 433, 434, and 436
each contains the amino acids Y, T, E, K, F, and Y, and the FcRn antagonist is directed to the target.
The above method, wherein the method is administered at least twice in 20 days.
(Configuration 3)
2. The method of configuration 2, wherein the FcRn antagonist is administered to the subject once every 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days.
(Configuration 4)
2. The method of configuration 2, wherein the FcRn antagonist is administered to the subject once every four days.
(Configuration 5)
2. The method of configuration 2, wherein the FcRn antagonist is administered to the subject once every 7 days.
(Configuration 6)
The FcRn antagonist is administered to the subject on day 20 at 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
The method of configuration 2 or 3, wherein the method is administered 13, 14, 15, 16, 17, 18, 19, or 20 times.
(Configuration 7)
A method of reducing serum levels of an Fc-containing agent in a subject, the method comprising administering to the subject an effective amount of an isolated FcRn antagonist comprising a variant Fc region or an FcRn-binding fragment thereof, the method comprising: Fc domains of EU positions 252, 254, 256, 433, 434, and 436
each contains the amino acids Y, T, E, K, F, and Y, and the FcRn antagonist is directed to the target.
The above method, wherein the method is administered once every 48 hours for 4 weeks.
(Configuration 8)
wherein said FcRn antagonist is administered to said subject at a dose of about 0.2 to about 200 mg/kg.
The method described in any one of items 2 to 6.
(Configuration 9)
The FcRn antagonist administers to the subject about 0.2, 1, 2, 3, 5, 10, 20, 25, 30, 50, 70
, 100, or 200 mg/kg.
(Configuration 10)
10. The method of any one of configurations 1-9, wherein said FcRn antagonist is administered to said subject at a dose of about 10 mg/kg.
(Configuration 11)
11. The method of any one of configurations 1-10, wherein said FcRn antagonist is administered to said subject at a dose of about 20 mg/kg.
(Configuration 12)
12. The method of any one of configurations 1-11, wherein said FcRn antagonist is administered to said subject at a dose of about 25 mg/kg.
(Configuration 13)
13. The method of any one of configurations 1-12, wherein the FcRn antagonist is administered intravenously.
(Configuration 14)
14. The method of any one of configurations 1-13, wherein the FcRn antagonist is administered subcutaneously.
(Configuration 15)
Configurations 1-14, wherein the FcRn antagonist is administered to the subject in two or more doses, wherein the first dose administered to the subject is administered intravenously and the subsequent one or more doses are administered subcutaneously.
The method described in any one of the following.
(Configuration 16)
16. The method of any one of configurations 1-15, wherein the FcRn antagonist does not include an antibody variable region.
(Configuration 17)
17. The method of any one of configurations 1-16, wherein the FcRn antagonist does not include a CH1 domain.
(Configuration 18)
18. The method of any one of configurations 1-17, wherein the FcRn antagonist does not contain free cysteine residues.
(Configuration 19)
19. The method according to any one of configurations 1 to 18, wherein the variant Fc region is an IgG Fc region.
(Configuration 20)
20. The method according to any one of configurations 1 to 19, wherein the variant Fc region is an IgG1 Fc region.
(Configuration 21)
21. The method according to any one of configurations 1 to 20, wherein the amino acid sequence of the Fc domain of the variant Fc region comprises the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3.
(Configuration 22)
22. The method according to any one of configurations 1 to 21, wherein the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO: 1.
(Configuration 23)
Configurations 1 to 22, wherein the FcRn antagonist consists of a variant Fc region, and the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3.
The method described in any one of the following.
(Configuration 24)
24. The method of any one of configurations 1-23, wherein the variant Fc region has increased affinity for Fcγ receptors compared to the affinity of the wild-type IgG1 Fc region for Fcγ receptors.
(Configuration 25)
25. The method of any one of configurations 1-24, wherein the variant Fc region has increased affinity for CD16a.
(Configuration 26)
26. The method of any one of configurations 1-25, wherein the Fc domain of the variant Fc region does not contain an N-linked glycan at EU position 297.
(Configuration 27)
The FcRn antagonist comprises a plurality of FcRn antagonist molecules, at least 50% (optionally at least 60, 70, 80, 90, 95, or 99%) of the plurality of FcRn antagonist molecules having an afucosylated N at EU position 297. - The method according to any one of configurations 1 to 25, comprising a variant Fc region or an FcRn-binding fragment thereof comprising a linked glycan.
(Configuration 28)
The FcRn antagonist comprises a plurality of FcRn antagonist molecules, at least 50% (optionally at least 60, 70, 80, 90, 95, or 99%) of the plurality of FcRn antagonist molecules having a bisecting GlcNac at EU position 297. Variant Fc containing N-linked glycans with
26. The method according to any one of configurations 1 to 25, comprising the region or FcRn-binding fragment thereof.
(Configuration 29)
29. The method of any one of configurations 1-28, wherein the variant Fc region is linked to a half-life extender.
(Configuration 30)
the half-life extender is polyethylene glycol or human serum albumin;
The method described in any one of configurations 1 to 29.
(Configuration 31)
31. The method according to any one of configurations 1 to 30, wherein the Fc-containing agent is an antibody or an immunoadhesin.
(Configuration 32)
32. The method according to any one of configurations 1 to 31, wherein the Fc-containing agent is a therapeutic agent or a diagnostic agent.
(Configuration 33)
33. The method according to any one of configurations 1 to 32, wherein the Fc-containing agent is a contrast agent.
(Configuration 34)
34. The method of any one of configurations 1-33, wherein the Fc-containing agent is an antibody-drug conjugate.
(Configuration 35)
35. The method of any one of configurations 1-34, wherein the Fc-containing agent is a pathogenic antibody.
(Configuration 36)
36. The method according to any one of configurations 1 to 35, wherein the Fc-containing agent is an autoantibody.
(Configuration 37)
37. The method of any one of structures 1-36, wherein said subject has an antibody-mediated disease or disorder, and wherein said disease or disorder is ameliorated by administering said FcRn antagonist to said subject.
(Configuration 38)
38. The method of structure 37, wherein the antibody-mediated disease or disorder is an autoimmune disease.
(Configuration 39)
The autoimmune disease is allogeneic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune Addison's disease, Alzheimer's disease, anti-neutrophil cytoplasmic autoantibodies (ANCA) , autoimmune diseases of the adrenal glands, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia. autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman syndrome, celiac disease-dermatitis, chronic fatigue immunodeficiency syndrome, chronic inflammatory demyelinating polyneuritis (C
IDP), Churg-Strauss syndrome, cicatricial pemphigoid, Crest syndrome, cold agglutinin disease,
Crohn's disease, dermatomyositis, dilated cardiomyopathy, discoid lupus erythematosus, epidermolysis bullosa acquired, essential mixed cryoglobulinemia, factor VIII deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Graves disease, Guillain-Barre, Goodpasture syndrome, graft-versus-host disease (GVHD)
, Hashimoto's thyroiditis, hemophilia A, idiopathic membranous neuropathy, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, IgM polyneuropathy, immune-mediated thrombocytopenia, juvenile onset Arthritis, Kawasaki disease, lichen planus, lichen sclerosus, lupus erythematosus, Meniere's disease, mixed connective tissue disease, mucosal pemphigoid, multiple sclerosis, type 1 diabetes, multifocal motor neuropathy (MMN), myasthenia gravis , paraneoplastic bullous pemphigoid, gestational pemphigoid, pemphigus vulgaris, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, relapsing polychondritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma Sjögren syndrome, solid organ transplant rejection, stiff man syndrome, systemic lupus erythematosus, Takayasu arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (SJS), temporal arteritis/giant cell syndrome arteritis,
selected from the group consisting of thrombotic thrombocytopenic purpura, ulcerative colitis, uveitis, dermatitis herpetiformis vasculitis, anti-neutrophil cytoplasmic antibody-associated vasculitis, vitiligo, and Wegener's granulomatosis ,
The method described in Configuration 38.
(Configuration 40)
38. The method of claim 37, wherein the disease or disorder is treatable using intravenous immunoglobulin (IVIG), plasmapheresis, and/or immunoadsorption.
(Configuration 41)
39. The method according to constitution 38, wherein the autoimmune disease is an autoimmune channelopathy.
(Configuration 42)
The channelopathies include autoimmune limbic encephalitis, epilepsy, neuromyelitis optica, Lambert-Eaton myasthenia syndrome, myasthenia gravis, anti-N-methyl-D-aspartate (NMDA) receptor encephalitis, anti-α -Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor encephalitis, Morvan syndrome, myotonia nervosa, pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS) ), and glycine receptor antibody-related disorders.
The method described in 41.
(Configuration 43)
38. The method of structure 37, wherein the antibody-mediated disease or disorder is hyperglobulinemia.
(Configuration 44)
44. The method of any one of configurations 1-43, wherein the FcRn antagonist is administered to the subject simultaneously or sequentially with an additional therapeutic agent.
(Configuration 45)
45. The method of clause 44, wherein the additional therapeutic agent is an anti-inflammatory drug.
(Configuration 46)
45. The method of configuration 44, wherein the additional therapeutic agent is a leukapheresis agent.
(Configuration 47)
46. The method of configuration 45, wherein the leukocyte depleting agent is a B cell depleting agent.
(Configuration 48)
48. The method according to configuration 47, wherein the B cell depletion agent is an antibody.
(Configuration 49)
The B cell depletion drug is CD10, CD19, CD20, CD21, CD22, CD23, CD24, CD37, CD53, CD70.
, CD72, CD74, CD75, CD77, CD79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85, or
49. The method of configuration 48, wherein the antibody specifically binds to CD86.
(Configuration 50)
The additional therapeutic agent may include rituximab, daclizumab, basiliximab, muronomab-CD3
, infliximab, adalimumab, omalizumab, efalizumab, natalizumab, tocilizumab, eculizumab, golimumab, canakinumab, ustekinumab, belimumab,
or a combination thereof, the method according to configuration 49.
(Configuration 51)
51. The method according to any one of configurations 1 to 50, wherein the subject is a human or a cynomolgus monkey.

Claims (27)

A pharmaceutical composition for use in a method of reducing serum levels of autoantibodies in a subject, comprising an FcRn antagonist consisting of a variant Fc region;
The amino acid sequence of at least one Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO: 1, 2 or 3 ,
Said FcRn antagonist is administered to said subject at a dose of 2 to 200 mg/kg, and said FcRn antagonist is administered to said subject twice every 20 days. 2. The pharmaceutical composition of claim 1, wherein the FcRn antagonist is administered intravenously to the subject. 2. The pharmaceutical composition of claim 1, wherein the FcRn antagonist is administered subcutaneously. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the amino acid sequence of at least one Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO:1. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the amino acid sequence of at least one Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO:2. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the amino acid sequence of at least one Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO:3. The pharmaceutical composition according to any one of claims 1 to 3, wherein the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO: 1, 2, or 3 . The pharmaceutical composition according to any one of claims 1 to 3 , wherein the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO:1. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO:2. The pharmaceutical composition according to any one of claims 1 to 3 , wherein the amino acid sequence of the Fc domain of the variant Fc region consists of the amino acid sequence set forth in SEQ ID NO:3. 11. The pharmaceutical composition of any one of claims 1 to 10 , wherein the FcRn antagonist is administered once every 7, 8, 9, or 10 days. A pharmaceutical composition according to any one of claims 1 to 11 , wherein the FcRn antagonist is administered once every 7 days. A pharmaceutical composition according to any one of claims 1 to 11 , wherein the FcRn antagonist is administered once every 10 days. 14. The pharmaceutical composition of any one of claims 1-13 , wherein the variant Fc region is linked to a half-life extender, the half-life extender being polyethylene glycol or human serum albumin. 15. The pharmaceutical composition of any one of claims 1-14 , wherein the subject has an antibody-mediated disease or disorder. The pharmaceutical composition according to any one of claims 1 to 10 , wherein the subject has an autoimmune disease. The autoimmune disease is allogeneic islet graft rejection, alopecia areata, ankylosing spondylitis, antiphospholipid antibody syndrome, autoimmune Addison's disease, Alzheimer's disease, anti-neutrophil cytoplasmic autoantibodies (ANCA) , autoimmune diseases of the adrenal glands, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune myocarditis, autoimmune neutropenia, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia. autoimmune urticaria, Behcet's disease, bullous pemphigoid, cardiomyopathy, Castleman's disease, celiac disease-dermatitis, chronic fatigue immunodeficiency syndrome, chronic inflammatory demyelinating polyneuritis (CIDP), churg - Strauss syndrome, cicatricial pemphigoid, Crest syndrome, cold agglutinin disease, Crohn's disease, dermatomyositis, dilated cardiomyopathy, discoid lupus erythematosus, epidermolysis bullosa acquired, mixed essential cryoglobulinemia, Factor VIII deficiency, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, Goodpasture syndrome, graft-versus-host disease (GVHD), Hashimoto's thyroiditis, hemophilia A, idiopathic membranes sexual neuropathy, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA neuropathy, IgM polyneuropathy, immune-mediated thrombocytopenia, juvenile arthritis, Kawasaki disease, lichen planus, lichen sclerosus, lupus erythematosus , Meniere's disease, mixed connective tissue disease, mucosal pemphigoid, multiple sclerosis, type 1 diabetes, multifocal motor neuropathy (MMN), myasthenia gravis, paraneoplastic bullous pemphigoid, gravida Pemphigus, pemphigus vulgaris, pemphigus foliaceus, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agamma Globulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, relapsing polychondritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, solid organ transplant rejection, stiff Mann syndrome, systemic lupus erythematosus, Takayasu arteritis, toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome (SJS), temporal arteritis/giant cell arteritis, thrombotic thrombocytopenic purpura, ulcerative 17. The pharmaceutical composition of claim 16 , selected from the group consisting of colitis, uveitis, herpeticoid vasculitis, anti-neutrophil cytoplasmic antibody-associated vasculitis, vitiligo, and Wegener's granulomatosis. 17. The pharmaceutical composition according to claim 16 , wherein the autoimmune disease is an autoimmune channelopathy. The channelopathies include autoimmune limbic encephalitis, epilepsy, neuromyelitis optica, Lambert-Eaton myasthenia syndrome, myasthenia gravis, anti-N-methyl-D-aspartate (NMDA) receptor encephalitis, anti-α -Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor encephalitis, Morvan syndrome, myotonia nervosa, pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS) ), and anti - glycine receptor antibody-related disorders. 20. The pharmaceutical composition of any one of claims 1-19 , wherein the FcRn antagonist is administered to the subject simultaneously or sequentially with an additional therapeutic agent. 21. The pharmaceutical composition of claim 20 , wherein the additional therapeutic agent is an anti-inflammatory agent or a leukapheresis agent. 22. The pharmaceutical composition according to claim 21 , wherein the leukocyte depleting agent is a B cell depleting agent. 23. The pharmaceutical composition of claim 22 , wherein the B cell depleting agent is an antibody. The B cell removing drug is CD10, CD19, CD20, CD21, CD22, CD23, CD24, CD37, CD53, CD70, CD72, CD74, CD75, CD77, CD79a, CD79b, CD80, CD81, CD82, CD83, CD84, CD85. 24. The pharmaceutical composition according to claim 23 , which is an antibody that specifically binds to CD86. 21. The additional therapeutic agent is rituximab, daclizumab, basiliximab, muromonab -CD3, infliximab, adalimumab, omalizumab, efalizumab, natalizumab, tocilizumab, eculizumab, golimumab, canakinumab, ustekinumab, belimumab, or a combination thereof. Pharmaceutical composition. 26. The pharmaceutical composition according to any one of claims 1 to 25 , wherein the subject is a human or a cynomolgus monkey. 27. The pharmaceutical composition according to any one of claims 1 to 26 , wherein the subject is a human.

Images